Mycoplasma pneumoniae IgM antibody colloid gold method detecting kit and preparation method thereof
A technology of mycoplasma pneumoniae and colloidal gold, which is applied in the direction of measuring devices, instruments, scientific instruments, etc., can solve the problems of low sensitivity, poor specificity of mycoplasma pneumoniae, and limited clinical application value, achieving high sensitivity, obvious effect, and easy operation Effect
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Embodiment 1
[0058] 1. Source of raw materials
[0059] The source and dosage of each raw material and diluent prepared by the Mycoplasma pneumoniae IgM antibody detection kit (colloidal gold method) are shown in Table 10 below:
[0060]
[0061] 2. Raw material requirements:
[0062] 2.1 Recombinant MP antigen
[0063] (1) Appearance: colorless transparent liquid;
[0064] (2) Concentration and purity requirements: the concentration is greater than 2mg / ml, determined by SDS-PAGE, and there is only one band under the condition of 10μl sample loading;
[0065] 2.2 Goat anti-mouse IgG antibody
[0066] (1) Appearance: colorless transparent liquid;
[0067] (2) Concentration requirements: concentration greater than 4mg / ml;
[0068] 2.3 Mouse anti-human IgM monoclonal antibody
[0069] (1) Appearance: colorless transparent liquid;
[0070] (2) Concentration and purity requirements: the concentration is greater than 2 mg / ml, detected by SDS-PAGE protein electrophoresis, and the he...
Embodiment 2
[0151] Kit preparation one
[0152] (1) Preparation of reaction membrane: Dilute the recombinant MP antigen with phosphate buffer solution to a coating concentration of 1.0mg / ml, dilute the goat anti-mouse IgG antibody to a coating concentration of 2.0mg / ml, and use a film dispenser to apply The two coating solutions were drawn onto the nitrocellulose membrane respectively, and the coated nitrocellulose membrane was dried and stored;
[0153] (2) Preparation of mouse anti-human IgM monoclonal antibody gold conjugate pad: Colloidal gold-labeled mouse anti-human IgM monoclonal antibody solution was prepared by coupling mouse anti-human IgM monoclonal antibody with a labeling concentration of 10 μg / ml to colloidal gold. Centrifuge at 12000r / min for 40min, discard the supernatant, add the colloidal gold conjugate diluent to 1 / 2 of the original volume, then soak the gold standard pad with 1.5ml / strip of the mixed solution to prepare MP-IgM gold conjugate pad , put the MP-IgM go...
Embodiment 3
[0157] Kit Preparation II
[0158] (1) Preparation of reaction membrane: Dilute the recombinant MP antigen with phosphate buffer solution to a coating concentration of 1.0mg / ml, dilute the goat anti-mouse IgG antibody to a coating concentration of 2.0mg / ml, and use a film dispenser to apply The two coating solutions were drawn onto the nitrocellulose membrane respectively, and the coated nitrocellulose membrane was dried and stored;
[0159] (2) Preparation of mouse anti-human IgM monoclonal antibody gold conjugate pad: Colloidal gold-labeled mouse anti-human IgM monoclonal antibody solution was prepared by coupling mouse anti-human IgM monoclonal antibody with a labeling concentration of 18 μg / ml to colloidal gold. Centrifuge at 12000r / min for 40min, discard the supernatant, add the colloidal gold conjugate diluent to 1 / 2 of the original volume, then soak the gold standard pad with 1.5ml / strip of the mixed solution to prepare MP-IgM gold conjugate pad , put the MP-IgM gol...
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